Revealing microbial community structures in large- and small-scale activated sludge systems by barcoded pyrosequencing of 16S rRNA gene.
Revealing microbial community structures in large- and small-scale activated sludge systems by barcoded pyrosequencing of 16S rRNA gene.
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DOI：
10.2166/wst.2012.428
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发表时间：
2012-09
期刊：
影响因子：
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通讯作者：
P. Ranasinghe;H. Satoh;M. Oshiki;K. Oshima;W. Suda;M. Hattori;T. Mino
中科院分区：
文献类型：
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作者：
P. Ranasinghe;H. Satoh;M. Oshiki;K. Oshima;W. Suda;M. Hattori;T. Mino
采用16S rRNA基因的条形码焦磷酸测序技术，研究了大型和小型污水处理厂活性污泥中细菌群的多样性。从12个污水处理厂收集活性污泥样本（3个小型和17个大型），以明确准确的分类和相对丰度。提取DNA，用4个碱基条形码27f/519r引物扩增。获得了454个Titanium （Roche）焦序，并通过Quantitative Insight Into Microbial Ecology （QIIME）进行了大约100,000个reads的分析。计算序列统计量，构建系统发育树和热图。计算结果解释了门和纲水平上的总微生物多样性，并进一步扩展到基于操作分类单元（OTU）的分类分配，以调查基于个体样本的群落分布。比较了大型和小型污水处理厂的微生物群落结构，确定了主要门（变形菌门和拟杆菌门）和纲（β变形菌门和拟杆菌门）。此外，还解释了科水平的分解，并说明了在其物种水平上发现的硝化菌科和放线菌门的差异。因此，焦磷酸测序方法提供了对活性污泥中微生物群落结构的高分辨率洞察，这可能是传统方法所忽视的。
The diversity of bacterial groups in activated sludge from large- and small-scale wastewater treatment plants was explored by barcoded pyrosequencing of 16S rRNA gene. Activated sludge samples (three small and 17 large scale) were collected from 12 wastewater treatment plants to clarify precise taxonomy and relative abundances. DNA was extracted, and amplified by 4 base barcoded 27f/519r primer set. The 454 Titanium (Roche) pyrosequences were obtained and analyses performed by Quantitative Insight Into Microbial Ecology (QIIME) with around 100,000 reads. Sequence statistics were computed, while constructing a phylogenetic tree and heatmap. Computed results explained total microbial diversity at phylum and class level and resolution was further extended to Operational Taxonomic Unit (OTU) based taxonomic assignment for investigating community distribution based on individual sample. Composition of sequence reads were compared and microbial community structures for large- and small-scale treatment plants were identified as major phyla (Proteobacteria and Bacteroidetes) and classes (Betaproteobacteria and Bacteroidetes). Also, family level breakdowns were explained and differences in family Nitrospiraceae and phylum Actinobacteria found at their species level were also illustrated. Thus, the pyrosequencing method provides high resolution insight into microbial community structures in activated sludge that might have been unnoticed with conventional approaches.